Volume : 13, Issue : 08, August – 2026
Title:
DEVELOPMENT AND VALIDATION OF A STABILITY-INDICATING UPLC METHOD FOR THE SIMULTANEOUS ESTIMATION OF BICTEGRAVIR, EMTRICITABINE AND TENOFOVIR ALAFENAMIDE IN BULK AND PHARMACEUTICAL DOSAGE FORMS.
Authors :
Dr. G. Thara*, Biradar Bindu
Abstract :
The present study was undertaken to develop and validate a simple, rapid, precise, accurate, robust, and stability-indicating Ultra Performance Liquid Chromatography (UPLC) method for the simultaneous estimation of Bictegravir, Emtricitabine, and Tenofovir Alafenamide in bulk drug substances and pharmaceutical dosage forms. The chromatographic separation was achieved using an ACQUITY BEH C18 column (100 × 2.1 mm, 1.7 µm) with an isocratic mobile phase consisting of 0.1% orthophosphoric acid buffer and acetonitrile (50:50, v/v) at a flow rate of 0.35 mL/min. Detection was carried out at 260 nm using a PDA detector. The developed method produced well-resolved peaks with satisfactory retention times and excellent chromatographic performance. Validation was performed according to ICH Q2(R2) guidelines by evaluating system suitability, specificity, linearity, accuracy, precision, robustness, sensitivity, and solution stability. The calibration curves exhibited excellent linearity with correlation coefficients greater than 0.999 for all analytes. Recovery values ranged between 99% and 101%, while precision studies showed percentage relative standard deviation values below 2%, confirming the reliability of the method. Forced degradation studies under acidic, alkaline, oxidative, thermal, photolytic, and neutral conditions demonstrated effective separation of degradation products from the analyte peaks, confirming the stability-indicating nature of the method. The proposed UPLC method is rapid, reproducible, economical, and suitable for routine quality control analysis, stability testing, and regulatory applications of combined antiretroviral pharmaceutical formulations.
Keywords: Bictegravir; Emtricitabine; Tenofovir Alafenamide; Stability-Indicating UPLC; Method Validation.
Cite This Article:
Please cite this article in press Dr. G. Thara et al., Development and Validation of a Stability-Indicating UPLS method for the Simultaneous Estimation of Bictegravir, Emtricitabine and Tenofovir Alafenamide in bulk and pharmaceutical dosage forms.,, Indo Am. J. P. Sci, 2026; 13(08).
REFERENCES:
1. Ray AS, Fordyce MW, Hitchcock MJM. Tenofovir alafenamide: A novel prodrug of tenofovir for the treatment of human immunodeficiency virus. Antiviral Res. 2016;125:63–70.
2. Markowitz M, Zolopa A, Squires K, Ruane P, Coakley D, Kearney B, et al. Phase I/II study of the pharmacokinetics, safety and antiretroviral activity of tenofovir alafenamide, a new prodrug of the HIV reverse transcriptase inhibitor tenofovir, in HIV-infected adults. J Antimicrob Chemother. 2014;69(5):1362–1369.
3. Valade E, Treluyer JM, Bouazza N, Ghosn J, Foissac F, Benaboud S, et al. Population pharmacokinetics of emtricitabine in HIV-1-infected adult patients. Antimicrob Agents Chemother. 2014;58(4):2256–2261.
4. Tsiang M, Jones GS, Goldsmith J, Mulato A, Hansen D, Kan E, et al. Antiviral activity of bictegravir (GS-9883), a novel potent HIV-1 integrase strand transfer inhibitor with an improved resistance profile. Antimicrob Agents Chemother. 2016;60(12):7086–7097.
5. Deeks ED. Bictegravir/Emtricitabine/Tenofovir Alafenamide: A review in HIV-1 infection. Drugs. 2018;78(17):1817–1828.
6. Tanuja A, Ganapaty S, Satya Narayana Murthy V. Development and validation of a stability-indicating RP-HPLC method for the simultaneous estimation of bictegravir, emtricitabine and tenofovir alafenamide fumarate. Turk J Pharm Sci. 2021;18(4):410–419.
7. Vamsi D, Sowjanya G. Validated method for the simultaneous determination of emtricitabine, bictegravir and tenofovir alafenamide in pharmaceutical dosage form using UPLC. Int J Pharm Sci Res. 2022;13(11):4536–4543.
8. Mehetre N, Singh I. Advanced, cutting-edge RP-HPLC methodology for the comprehensive quantitative analysis of assay of bictegravir, emtricitabine and tenofovir alafenamide in co-formulated pharmaceutical products. Int J Pharm Qual Assur. 2024;15(3):1297–1302.
9. Tej Kumar K, Suryakala D. RP-HPLC method development and validation for the estimation of emtricitabine, bictegravir and tenofovir alafenamide in bulk and pharmaceutical dosage form. J Taibah Univ Sci. 2019;13(1):1137–1146.
10. Raja Reddy A, Devicharan R, Rama Rao T. Analytical RP-HPLC method development and validation for simultaneous estimation of emtricitabine, tenofovir and bictegravir. Res J Pharm Technol. 2023;16(8):3722–3726.
11. Tiris G, Genc AA, Oven E, Erk N. A green approach for the analysis of emtricitabine, bictegravir and tenofovir in a pharmaceutical preparation using novel HPLC and spectrophotometric methods. Biomed Chromatogr. 2023;37(10):e5712.
12. Vijay Swaroop Singh G, Divakar TE. A novel stability indicating RP-HPLC method development and validation for simultaneous estimation of bictegravir, emtricitabine and tenofovir in pure and fixed dose combination. Asian J Org Med Chem. 2019;4(1):7–13.
13. Asma Begum, Ayesha Begum K, Sandhya P. Analytical method development and validation of emtricitabine, bictegravir and tenofovir alafenamide by RP-HPLC. Int J Pharm Ind Res. 2020;10(4):179–186.
14. International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH). ICH Harmonised Tripartite Guideline Q2(R1): Validation of Analytical Procedures: Text and Methodology. Geneva: ICH; 2005.
15. International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH). ICH Harmonised Tripartite Guideline Q1A(R2): Stability Testing of New Drug Substances and Products. Geneva: ICH; 2003.